Preparation method of lysophosphatidylcholine

Abstract

The invention belongs to the technical field of chemical engineering, and provides a preparation method of lysophosphatidylcholine, which comprises the following steps: mixing glycerophosphorylcholine, dialkyl tin oxide and an organic solvent, and carrying out complex reaction to obtain a complex intermediate; mixing the system after the complexation reaction with fatty acid acyl chloride and an acid-binding agent, carrying out acylation reaction, adding water for quenching after the reaction is completed, filtering, and collecting filtrate; mixing the filtrate with an extraction agent for extraction, collecting a raffinate phase obtained by extraction, and carrying out vacuum concentration to obtain a lysophosphatidylcholine crude product; the lysophosphatidylcholine crude product is subjected to recrystallization, and lysophosphatidylcholine is obtained; the solvent for recrystallization is short-chain alcohol, the initial temperature of recrystallization is 45-75 DEG C, the crystallization temperature is 0-30 DEG C, and the cooling rate is 0.5-5 DEG C / min. The preparation method provided by the invention is short in process route, simple in product purification, relatively high in product purity and suitable for large-scale production.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical engineering, and in particular relates to a method for preparing lysophosphatidylcholine. Background Art

[0002] Lysophosphatidylcholine (LysoPC) is composed of a hydrophilic phosphatidylcholine head group and a hydrophobic fatty acid chain. It has surface activity and physiological activity and is widely used in the fields of medicine, food, and cosmetics. Lysophosphatidylcholine is one of the main ingredients used to prepare dosage forms such as nanoparticles and liposomes. Due to its unique biological activity and affinity for cell membranes, it can improve the bioavailability of drugs and help drugs effectively penetrate cell membranes. It plays an important role in the delivery of anticancer drugs and vaccines. In addition, lysophosphatidylcholine, as a biomarker, has been used in the diagnosis and disease monitoring of diseases such as atherosclerosis and diabetes. In the food industry, lysophosphatidylcholine is an excellent emulsifier that can stabilize fat in emulsified foods, improve food texture and taste, and extend shelf life. It is often used in baking, chocolate manufacturing, and dairy processing. In addition, lysophosphatidylcholine is widely used as an ingredient in functional foods or nutritional supplements because of its potential to improve lipid metabolism and anti-inflammatory effects. It is often used to assist in the treatment of fatty liver, abnormal cholesterol, and other problems. In the cosmetics industry, lysophosphatidylcholine can help improve skin barrier function, reduce water loss, and has a certain anti-inflammatory effect. Therefore, it is particularly important to prepare high-purity lysophosphatidylcholine.

[0003] The existing preparation method of lysophosphatidylcholine is mainly enzymatic hydrolysis, in which phosphatidylcholine is hydrolyzed by phospholipase A1 or A2 to remove a fatty acid chain to obtain lysophosphatidylcholine containing a fatty acid chain. However, due to the relatively high price of phospholipase and the relatively harsh reaction conditions, its industrial application is limited. At the same time, since the natural phosphatidylcholine raw material is a mixture containing different acyl groups, it is difficult to obtain lysophosphatidylcholine containing a single acyl group by enzymatic hydrolysis.

[0004] Chemical synthesis is an effective method for preparing high-purity lysophosphatidylcholine with a single chemical structure. The existing chemical synthesis method for preparing lysophosphatidylcholine cannot achieve regioselective esterification, has a long synthesis route, and is complex in reaction, and is not suitable for industrial production. Summary of the invention

[0005] In view of this, the present invention provides a method for preparing lysophosphatidylcholine. The preparation method provided by the present invention is simple and easy to operate, suitable for industrial production, and the lysophosphatidylcholine prepared according to the preparation method provided by the present invention has high purity and high yield.

[0006] In order to solve the above technical problems, the present invention provides a method for preparing lysophosphatidylcholine, comprising the following steps:

[0007] Glycerolphosphocholine, dialkyltin oxide and an organic solvent are mixed and subjected to a complex reaction to obtain a complex intermediate;

[0008] The system after the complex reaction is mixed with fatty acid chloride and an acid binding agent to carry out an acylation reaction, and after the reaction is completed, water is added to quench the reaction, and the filtrate is filtered to collect the filtrate;

[0009] The filtrate and the extractant are mixed for extraction, and the raffinate phase obtained by the extraction is collected and concentrated under reduced pressure to obtain a crude lysophosphatidylcholine product;

[0010] The crude lysophosphatidylcholine is recrystallized to obtain lysophosphatidylcholine; the recrystallization solvent is a short-chain alcohol, the starting temperature of the recrystallization is 45-75° C., the crystallization temperature is 0-30° C., and the cooling rate is 0.05-5° C. / min.

[0011] Preferably, the dialkyltin oxide includes dimethyltin oxide, diethyltin oxide or dibutyltin oxide;

[0012] The molar ratio of the glycerophosphocholine to the dialkyltin oxide is 1:(1-2).

[0013] Preferably, the organic solvent comprises an alcohol having 1 to 4 carbon atoms.

[0014] Preferably, the temperature of the complexation reaction is 50-80° C. and the time is 0.5-8 h.

[0015] Preferably, the carbon number of the fatty acid chloride is 8 to 24;

[0016] The mass ratio of the glycerophosphocholine to the volume ratio of the fatty acid chloride is 5 g: (2.5-9.8) mL.

[0017] Preferably, the acid binding agent comprises triethylamine, pyridine or 4-dimethylaminopyridine.

[0018] Preferably, the acylation reaction is carried out at a temperature of 20 to 60° C. and for a time of 0.5 to 12 h.

[0019] Preferably, the extractant is an alkane having 6 to 10 carbon atoms.

[0020] Preferably, the extraction is performed 2 to 4 times.

[0021] Preferably, the short-chain alcohol is an alcohol having 2 to 4 carbon atoms.

[0022] The present invention provides a method for preparing lysophosphatidylcholine, comprising the following steps: mixing glycerophosphocholine, dialkyltin oxide and an organic solvent and then performing a complex reaction to obtain a complex intermediate; mixing the system after the complex reaction with fatty acid chloride and an acid binding agent to perform an acylation reaction, adding water to quench the reaction after completion, filtering, and collecting the filtrate; mixing the filtrate with an extractant for extraction, collecting the raffinate phase obtained by the extraction and concentrating under reduced pressure to obtain a crude lysophosphatidylcholine; recrystallizing the crude lysophosphatidylcholine to obtain lysophosphatidylcholine; the recrystallization solvent is a short-chain alcohol, the starting temperature of the recrystallization is 45 to 75°C, the crystallization temperature is 0 to 30°C, and the cooling rate is 0.05 to 5°C / min. The preparation method provided by the present invention has a short process route, mild conditions, simple product purification, and is suitable for large-scale production; the lysophosphatidylcholine prepared according to the preparation method provided by the present invention has a high purity and a high yield. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The palm lysophosphatidylcholine prepared in Example 5 1 H-NMR spectrum;

[0024] Figure 2 This is the mass spectrum of palm lysophosphatidylcholine prepared in Example 5. DETAILED DESCRIPTION

[0025] The present invention provides a method for preparing lysophosphatidylcholine, comprising the following steps:

[0026] Glycerolphosphocholine, dialkyltin oxide and an organic solvent are mixed and subjected to a complex reaction to obtain a complex intermediate;

[0027] The system after the complex reaction is mixed with fatty acid chloride and an acid binding agent to carry out an acylation reaction, and after the reaction is completed, water is added to quench the reaction, and the filtrate is filtered to collect the filtrate;

[0028] The filtrate and the extractant are mixed for extraction, and the raffinate phase obtained by the extraction is collected and concentrated under reduced pressure to obtain a crude lysophosphatidylcholine product;

[0029] The crude lysophosphatidylcholine is recrystallized to obtain lysophosphatidylcholine; the recrystallization solvent is a short-chain alcohol, the starting temperature of the recrystallization is 45-75° C., the crystallization temperature is 0-30° C., and the cooling rate is 0.05-5° C. / min.

[0030] The present invention mixes glycerophosphocholine, dialkyl tin oxide and an organic solvent and then performs a complex reaction to obtain a complex intermediate. As a specific embodiment of the present invention, the dialkyl tin oxide may include dimethyl tin oxide, diethyl tin oxide or dibutyl tin oxide; the molar ratio of the glycerophosphocholine and the dialkyl tin oxide may be 1:(1-2), and may be specifically 1:1, 1:1.1, 1:1.2, 1:1.3 or 1:1.4. When the molar ratio of glycerophosphocholine and dialkyl tin oxide is too low or too high, it will cause an increase in by-products. The ingredients prepared according to the above molar ratio can ensure that fewer by-products are generated and improve the purity of the product.

[0031] As a specific embodiment of the present invention, the organic solvent may include an alcohol having 1 to 4 carbon atoms, specifically methanol, ethanol, n-propanol or isopropanol. Before mixing, the organic solvent may be subjected to a water removal treatment to improve the raw material conversion rate and reduce the generation of by-products. The present invention has no special requirements for the water removal method, and the conventional method in the art can be used. If the water content in the organic solvent is high, the raw material conversion rate will be reduced and the by-products will increase.

[0032] As a specific embodiment of the present invention, the temperature of the complex reaction can be 50-80° C., preferably 70-80° C., specifically 70° C., 75° C. or 80° C. If the reaction temperature is too low, the reaction time is long and the raw material conversion rate is low; if the reaction temperature is too high, side reactions are likely to occur. The present invention limits the temperature of the complex reaction to the above range to facilitate obtaining a product of higher purity.

[0033] As a specific embodiment of the present invention, the time of the complex reaction can be 0.5 to 8 hours, preferably 1 to 3 hours, and can be specifically 1 hour, 2 hours or 3 hours. If the reaction time is too short, the reaction will be incomplete and the raw material conversion rate will be low; if the reaction time is too long, the intermediate product may decompose, resulting in a reduced yield. The present invention limits the time of the complex reaction to the above range to improve the yield of the product. In the present invention, the equation of the complex reaction is shown in formula a:

[0034]

[0035] After the complex reaction, the present invention will mix the system after the complex reaction with fatty acid chloride and an acid binding agent, perform an acylation reaction, add water to quench the reaction after completion, filter, and collect the filtrate. As a specific embodiment of the present invention, the carbon number of the fatty acid chloride can be 8 to 24, or 8 to 20, and can be specifically octanoyl chloride, lauroyl chloride, pentadecanoyl chloride, stearyl chloride or palmitoyl chloride; the mass of the glycerophosphocholine and the volume ratio of the fatty acid chloride can be 5g: (2.5 to 9.8) mL, and can be specifically 5g: 5.2 mL, 5g: 6.6 mL, 5g: 7.2 mL, 5g: 8.4 mL, 5g: 9 mL or 5g: 9.8 mL. When the molar ratio of the two is too low or too high, both will cause an increase in by-products, increasing the difficulty of subsequent separation and purification.

[0036] As a specific embodiment of the present invention, the acid binding agent may include triethylamine, pyridine or 4-dimethylaminopyridine.

[0037] As a specific embodiment of the present invention, the temperature of the acylation reaction can be 20-60°C, or 30-50°C, specifically 30°C, 40°C or 50°C; the time of the acylation reaction can be 0.5-12h, or 1-4h, specifically 1h, 2h, 3h or 4h. When the acylation reaction temperature is too low or the reaction time is too short, the reaction will be incomplete and the raw material conversion rate will be low; and when the acylation reaction temperature is too high or the reaction time is too long, the side reaction products will increase. The present invention limits the temperature and time of the acylation reaction within the above range to improve the yield and purity of the product.

[0038] In the present invention, the equation of the acylation reaction is shown in formula b:

[0039]

[0040] In formula b, n can be 7 to 23, and can be specifically 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 or 23.

[0041] As a specific embodiment of the present invention, the water used for quenching after the reaction is completed can be ultrapure water, and the volume ratio of the water to the acylation reaction system can be 1:1; the process of adding water is carried out under stirring conditions, and the stirring time can be 0.5 to 2 hours, specifically 0.5 hours, 1 hour, 1.5 hours or 2 hours.

[0042] The present invention has no special requirements for the filtration, and conventional methods in the art may be used.

[0043] After obtaining the filtrate, the present invention mixes the filtrate and the extractant for extraction, collects the raffinate phase obtained by the extraction, and concentrates under reduced pressure to obtain a crude lysophosphatidylcholine product. As a specific embodiment of the present invention, the extractant can be an alkane with a carbon number of 6 to 10, specifically n-hexane, n-heptane or n-octane; the number of extractions can be 2 to 4 times, specifically 3 times. The present invention can remove fat-soluble impurities by extraction, and the raffinate phase mainly includes lysophosphatidylcholine, and a small amount of unreacted glycerophosphocholine and other water-soluble impurities.

[0044] The present invention has no special requirements for the reduced pressure concentration, and the conventional method in the art can be used.

[0045] After obtaining the crude lysophosphatidylcholine, the present invention recrystallizes the crude lysophosphatidylcholine to obtain lysophosphatidylcholine. As a specific embodiment of the present invention, the recrystallization solvent can be a short-chain alcohol, the short-chain alcohol can be an alcohol with 2 to 4 carbon atoms, and the short-chain alcohol can be anhydrous ethanol, n-butanol or isopropanol; the starting temperature of the recrystallization can be 45 to 75°C, preferably 50 to 70°C, and can be specifically 50°C, 55°C, 60°C, 65°C, or 70°C; the crystallization temperature can be 0 to 30°C, and can be specifically 0°C, 5°C, 10°C, 15°C or 25°C; the cooling rate can be 0.05 to 5°C / min, and can be specifically 0.05°C / min, 0.1°C / min, 0.15°C / min, 0.5°C / min, 1°C / min or 2°C / min. If the starting temperature is too low and / or the crystallization temperature is too high, the yield may be reduced, while if the starting temperature is too high and / or the crystallization temperature is too low, the product purity may be reduced. If the cooling rate during the crystallization is too slow or too fast, the purity of the crystallized product will be affected. The present invention limits the recrystallization to the above conditions to improve the purity and yield of the product.

[0046] In order to further illustrate the present invention, the technical solutions provided by the present invention are described in detail below in conjunction with embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0047] Example 1

[0048] 5 g of glycerophosphocholine, 5.8 g of dibutyltin oxide and 400 mL of purified methanol after dehydration were mixed and stirred at 80° C. for 1 h to carry out complexation reaction to obtain a reaction product containing a complex intermediate;

[0049] After the complex reaction system was cooled to room temperature (30°C), 4.3 mL of triethylamine and 5.2 mL of octanoyl chloride were added in sequence, and the acylation reaction was carried out at 30°C with stirring for 4 h, and then cooled to room temperature (20°C), 400 mL of ultrapure water was added and stirred for 1 h to quench the acylation reaction, and then filtered to collect the filtrate;

[0050] Add 400 mL of n-hexane to the filtrate for extraction, repeat the extraction three times, retain the raffinate phase, and concentrate the collected raffinate phase under reduced pressure to obtain a crude product of octanoyl lysophosphatidylcholine;

[0051] The crude octanoyl lysophosphatidylcholine was added with 30 mL of anhydrous ethanol, heated to 50° C. for heat dissolution, slowly cooled to 0° C. at a cooling rate of 0.1° C. / min, stirred for crystallization, and the obtained solid was vacuum dried after filtration to obtain 6.0 g of pure octanoyl lysophosphatidylcholine.

[0052] Example 2

[0053] 5 g of glycerophosphocholine, 5.8 g of dibutyltin oxide and 400 mL of purified methanol after dehydration were mixed and stirred at 75° C. for 2 h to carry out complexation reaction to obtain a reaction product containing a complex intermediate;

[0054] After the complex reaction system was cooled to room temperature (20°C), 4 mL of triethylamine and 6.6 mL of lauroyl chloride were added in sequence, and the mixture was stirred at 40°C for acylation reaction for 3 h, then cooled to room temperature (20°C), 400 mL of ultrapure water was added and stirred for 1 h to quench the acylation reaction, and then filtered to collect the filtrate;

[0055] Add 400 mL of n-hexane to the filtrate for extraction, repeat the extraction three times, retain the raffinate phase, and concentrate the collected raffinate phase under reduced pressure to obtain a crude product of lauroyl lysophosphatidylcholine;

[0056] The crude lauroyl lysophosphatidylcholine was added to 40 mL of anhydrous ethanol, heated to 55°C for hot dissolution, slowly cooled to 5°C at a cooling rate of 0.15°C / min, stirred for crystallization, and the obtained solid was vacuum dried after filtration to obtain 6.9 g of pure lauroyl lysophosphatidylcholine.

[0057] Example 3

[0058] 5 g of glycerophosphocholine, 5.8 g of dibutyltin oxide and 400 mL of purified methanol after dehydration were mixed and stirred at 75° C. for 2 h to carry out complexation reaction to obtain a reaction product containing a complex intermediate;

[0059] After the complex reaction system was cooled to room temperature (20°C), 4 mL of pyridine and 6.6 mL of lauroyl chloride were added in sequence, and the acylation reaction was carried out at 40°C with stirring for 3 h. After that, the system was cooled to room temperature (20°C), 400 mL of ultrapure water was added and stirred for 1 h to quench the acylation reaction, and then filtered to collect the filtrate;

[0060] Add 400 mL of n-hexane to the filtrate for extraction, repeat the extraction three times, retain the raffinate phase, and concentrate the collected raffinate phase under reduced pressure to obtain a crude product of lauroyl lysophosphatidylcholine;

[0061] The crude lauroyl lysophosphatidylcholine was added to 40 mL of anhydrous ethanol, heated to 55°C for hot dissolution, slowly cooled to 5°C at a cooling rate of 0.5°C / min, stirred for crystallization, and the obtained solid was vacuum dried after filtration to obtain 6.3 g of pure lauroyl lysophosphatidylcholine.

[0062] Example 4

[0063] 5 g of glycerophosphocholine, 6.3 g of dibutyltin oxide and 400 mL of purified ethanol after dehydration were mixed and stirred at 70° C. for 3 h to carry out complex reaction to obtain a complex intermediate;

[0064] After the complex reaction system was cooled to room temperature (20°C), 5.4 mL of triethylamine and 9.8 mL of pentadecanoyl chloride were added in sequence, and the mixture was stirred at 50°C for 1 h for acylation reaction. After that, the mixture was cooled to room temperature (20°C), 400 mL of ultrapure water was added and stirred for 30 min to quench the acylation reaction, and then filtered to collect the filtrate;

[0065] Add 400 mL of n-heptane to the filtrate for extraction, repeat the extraction three times, retain the raffinate phase, and concentrate the collected raffinate phase under reduced pressure to obtain a crude product of pentadecanoyl lysophosphatidylcholine;

[0066] The crude pentadecanoyl lysophosphatidylcholine was added with 40 mL of isopropanol, heated to 70° C. for heat dissolution, slowly cooled to 15° C. at a cooling rate of 0.05° C. / min, stirred for crystallization, and the obtained solid was vacuum dried after filtration to obtain 8.2 g of pure pentadecanoyl lysophosphatidylcholine.

[0067] Example 5

[0068] 5 g of glycerophosphocholine, 4.8 g of dibutyltin oxide and 400 mL of purified ethanol after dehydration were mixed and stirred at 70° C. for 3 h to carry out complexation reaction to obtain a reaction product containing a complex intermediate;

[0069] After the complex reaction system was cooled to room temperature (20°C), 3.2 mL of triethylamine and 7.2 mL of palmitoyl chloride were added in sequence, and the mixture was stirred at 40°C for acylation reaction for 3 h, then cooled to room temperature (20°C), 400 mL of ultrapure water was added and stirred for 30 min to quench the acylation reaction, and then filtered to collect the filtrate;

[0070] Add 400 mL of n-heptane to the filtrate for extraction, repeat the extraction three times, retain the raffinate phase, and concentrate the collected raffinate phase under reduced pressure to obtain crude palm lysophosphatidylcholine;

[0071] The crude palm lysophosphatidylcholine was added to 40 mL of isopropanol, heated to 65°C for hot dissolution, slowly cooled to 10°C at a cooling rate of 0.5°C / min, stirred for crystallization, and the obtained solid was vacuum dried after filtration to obtain 8.2 g of pure palm lysophosphatidylcholine.

[0072] Example 6

[0073] 5 g of glycerophosphocholine, 7.2 g of dibutyltin oxide and 400 mL of purified isopropanol after dehydration were mixed and stirred at 80° C. for 1 h to carry out complex reaction to obtain a complex intermediate;

[0074] After the complex reaction system was cooled to room temperature (20°C), 4 mL of triethylamine and 8.4 mL of stearoyl chloride were added in sequence, and the acylation reaction was carried out at 40°C with stirring for 4 h. After that, the system was cooled to room temperature (20°C), 400 mL of ultrapure water was added and stirred for 30 min to quench the acylation reaction, and then filtered to collect the filtrate;

[0075] Add 400 mL of n-octane to the filtrate for extraction, repeat the extraction three times, retain the raffinate phase, and concentrate the collected raffinate phase under reduced pressure to obtain a crude stearoyl lysophosphatidylcholine product;

[0076] The crude stearoyl lysophosphatidylcholine was added to 50 mL of n-butanol, heated to 70°C for hot dissolution, slowly cooled to 25°C at a cooling rate of 0.5°C / min, stirred for crystallization, and the obtained solid was vacuum dried after filtration to obtain 8.5 g of pure stearoyl lysophosphatidylcholine.

[0077] Example 7

[0078] 5 g of glycerophosphocholine, 5.3 g of dibutyltin oxide and 400 mL of isopropanol were mixed and stirred at 80° C. for 1 h to carry out complex reaction to obtain a complex intermediate;

[0079] After the complex reaction system was cooled to room temperature (20°C), 4 mL of triethylamine and 9 mL of palmitoyl chloride were added in sequence, and the mixture was stirred at 50°C for 2 h for acylation reaction. After that, the mixture was cooled to room temperature (20°C), 400 mL of ultrapure water was added and stirred for 1 h to quench the acylation reaction, and then filtered to collect the filtrate;

[0080] Add 400 mL of n-heptane to the filtrate for extraction, repeat the extraction three times, retain the raffinate phase, and concentrate the collected raffinate phase under reduced pressure to obtain crude palmitoyl lysophosphatidylcholine;

[0081] The crude palmitoyl lysophosphatidylcholine was added with 40 mL of isopropanol, heated to 60° C. for heat dissolution, slowly cooled to 10° C. at a cooling rate of 2° C. / min, stirred for crystallization, and the obtained solid was vacuum dried after filtration to obtain 6.7 g of pure palmitoyl lysophosphatidylcholine.

[0082] Example 8

[0083] 5 g of glycerophosphocholine, 5.3 g of dimethyltin oxide and 400 mL of isopropanol were mixed and stirred at 80° C. for 1 h to carry out complex reaction to obtain a complex intermediate;

[0084] After the complex reaction system was cooled to room temperature (20°C), 4 mL of triethylamine and 9 mL of palmitoyl chloride were added in sequence, and the mixture was stirred at 50°C for 2 h for acylation reaction. After that, the mixture was cooled to room temperature (20°C), 400 mL of ultrapure water was added and stirred for 1 h to quench the acylation reaction, and then filtered to collect the filtrate;

[0085] Add 400 mL of n-heptane to the filtrate for extraction, repeat the extraction three times, retain the raffinate phase, and concentrate the collected raffinate phase under reduced pressure to obtain crude palmitoyl lysophosphatidylcholine;

[0086] The crude palmitoyl lysophosphatidylcholine was added with 40 mL of isopropanol, heated to 60° C. for heat dissolution, slowly cooled to 10° C. at a cooling rate of 0.5° C. / min, stirred for crystallization, and the obtained solid was vacuum dried after filtration to obtain 6.1 g of pure palmitoyl lysophosphatidylcholine.

[0087] Comparative Example 1

[0088] Acyl lysophosphatidylcholine was prepared according to Example 5, except that the hot melting temperature during the recrystallization process was 65° C., the cooling rate was 10° C. / min, and the crystallization temperature was 10° C. 8.6 g of palm lysophosphatidylcholine product was obtained.

[0089] Comparative Example 2

[0090] Acyl lysophosphatidylcholine was prepared according to Example 5, except that the hot melting temperature during the recrystallization process was 55° C., the cooling rate was 0.5° C. / min, and the crystallization temperature was 40° C. 6.3 g of palm lysophosphatidylcholine product was obtained.

[0091] Comparative Example 3

[0092] Acyl lysophosphatidylcholine was prepared according to Example 5, except that the hot melting temperature during the recrystallization process was 55° C., the cooling rate was 10° C. / min, and the crystallization temperature was 20° C. 7.1 g of palm lysophosphatidylcholine product was obtained.

[0093] A high performance liquid chromatography (HPLC) method was established to quantitatively analyze the purity of the lysophosphatidylcholine product, and the results are listed in Table 1. HPLC uses a charged aerosol detector (CAD), and the specific analysis method is as follows: The chromatographic column is a reverse phase column ACQUITUPLCBEH C 18 (100mm×2.1mm ID, 1.7μm), column temperature was 45℃. Mobile phase A was water, mobile phase B was isopropanol:acetonitrile = 1:1 (v:v), mobile phase A: mobile phase B = 48:52 (v:v), isocratic elution, flow rate was 0.2mL / min, injection volume was 2μL. The dilution solvent was n-hexane: isopropanol: water = 23:23:4 (v:v:v), concentration was 1mg / mL.

[0094] The yields of lysophosphatidylcholine prepared in Examples 1 to 8 and Comparative Examples 1 to 3 were calculated according to Formula 1, and the results are listed in Table 1.

[0095]

[0096] Wherein, GPC is glycerophosphocholine, m(GPC) is the GPC feed mass, and the molecular weight M(GPC) of GPC is 257.22 g / mol.

[0097] The HPLC purity of the lysophosphatidylcholine prepared in Examples 1 to 8 and Comparative Examples 1 to 3 was calculated according to the HPLC spectra according to Formula 2, and the results are listed in Table 1.

[0098] HPLC purity (%) = lysophosphatidylcholine peak area / sum of product component peak areas × 100% Formula 2.

[0099] Table 1 Yield and HPLC purity of lysophosphatidylcholine prepared in Examples 1 to 8 and Comparative Examples 1 to 3

[0100] Example Yield (%) HPLC purity (%) Example 1 79.2 98.3 Example 2 80.1 99.1 Example 3 78.0 98.1 Example 4 86.4 98.6 Example 5 83.7 98.4 Example 6 82.4 98.7 Example 7 83.8 91.7 Example 8 79.2 95.2 Comparative Example 1 74.9 84.2 Comparative Example 2 33.4 90.1 Comparative Example 3 68.6 85.7

[0101] As can be seen from Table 1, the lysophosphatidylcholine prepared according to the preparation method provided by the present invention has a higher yield and purity. By comparing the results of the examples and comparative examples, it can be seen that only by recrystallization under the conditions specified in the present invention can lysophosphatidylcholine with higher purity and higher recovery be prepared.

[0102] The product structure and purity were further confirmed by H NMR and mass spectrometry. 1 H-NMR and mass spectra are shown in Figure 1 and Figure 2 Described by Figure 1 and Figure 2It can be seen that the prepared product is palmitoyl lysophosphatidylcholine without obvious impurities.

[0103] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing lysophosphatidylcholine, characterized in that: The following steps are involved: Glycerolphosphocholine, dialkyltin oxide and an organic solvent are mixed and subjected to a complex reaction to obtain a complex intermediate; The system after the complex reaction is mixed with fatty acid chloride and an acid binding agent to carry out an acylation reaction, and after the reaction is completed, water is added to quench the reaction, and the filtrate is filtered to collect the filtrate; The filtrate and the extractant are mixed for extraction, and the raffinate phase obtained by the extraction is collected and concentrated under reduced pressure to obtain a crude lysophosphatidylcholine product; The crude lysophosphatidylcholine is recrystallized to obtain lysophosphatidylcholine; the recrystallization solvent is a short-chain alcohol, the starting temperature of the recrystallization is 45-75° C., the crystallization temperature is 0-30° C., and the cooling rate is 0.05-5° C. / min.

2. The preparation method according to claim 1, characterized in that: The dialkyltin oxide includes dimethyltin oxide, diethyltin oxide or dibutyltin oxide; The molar ratio of the glycerophosphocholine to the dialkyltin oxide is 1:(1-2).

3. The preparation method according to claim 1, characterized in that: The organic solvent includes alcohol having 1 to 4 carbon atoms.

4. The preparation method according to any one of claims 1 to 3, characterized in that: The temperature of the complex reaction is 50-80° C., and the time is 0.5-8 hours.

5. The preparation method according to claim 1, characterized in that: The carbon number of the fatty acid chloride is 8 to 24; The mass ratio of the glycerophosphocholine to the volume ratio of the fatty acid chloride is 5 g: (2.5-9.8) mL.

6. The preparation method according to claim 1, characterized in that: The acid binding agent includes triethylamine, pyridine or 4-dimethylaminopyridine.

7. The preparation method according to claim 1, 5 or 6, characterized in that: The temperature of the acylation reaction is 20-60° C., and the time is 0.5-12 h.

8. The preparation method according to claim 1, characterized in that: The extractant is an alkane having 6 to 10 carbon atoms.

9. The preparation method according to claim 8, characterized in that: The extraction is performed 2 to 4 times.

10. The preparation method according to claim 1, characterized in that: The short-chain alcohol is an alcohol having 2 to 4 carbon atoms.

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